KR102043250B1 - Microelectrode array chip capable of photothermal stimulation and control method of nerve cell using the same - Google Patents

Abstract

The present invention relates to a microelectrode chip and a method for controlling neuronal activity using the same, the microelectrode chip comprising: a substrate; An electrode pad disposed on a portion of the substrate; An electrode disposed on a portion of the substrate and connected to the electrode pad; An insulating layer disposed on a portion of the substrate where the electrode is not disposed; And a metal thin film disposed on the insulating layer.

Description

MICROELECTRODE ARRAY CHIP CAPABLE OF PHOTOTHERMAL STIMULATION AND CONTROL METHOD OF NERVE CELL USING THE SAME}

The present invention relates to a microelectrode chip utilizing the photothermal effect and a method for controlling neuronal activity using the same.

Regulating neuronal activity is essential for neurophysiological research and alleviation of clinical disease. To this end, microelectrode chips, which are formed by culturing neurons and forming networks, are controlled using various methods such as electricity and drugs to enhance or restore the function of the nervous system. In particular, the use of microelectrodes enables the measurement of microelectrical signals of neurons or electrical stimulation using microcurrents, so that the connection of neural networks can also be analyzed.

The electrical stimulation using the microelectrode is very effective in inducing excitatory stimulation to nerve cells, but has limitations in space-time resolution and difficult implementation of inhibitory stimulation.

As a new alternative technology to replace this, stimulation and photogenetics using infrared laser have been studied among non-invasive and highly efficient optical methods. Photostimulation can be achieved by expressing the photosensitive protein on the cell membrane mainly by genetic engineering through photogenetic techniques and using this to implement excitatory / inhibitory stimulation.

Photogenetics is a technology that artificially expresses ion channels that are specifically opened and closed by light in neurons, and then creates or eliminates cell action potentials using light in the visible region. However, photogenetics has a disadvantage in that it is difficult to apply in the clinic because genetic variation of neurons is required.

Accordingly, there is a need for an alternative technique to avoid this. In this regard, recently, photo-stimulation technology using the photothermal effect has been reported as an alternative technology for maintaining the advantages of photo-stimulation without genetic manipulation. In particular, when fabricating a microelectrode chip capable of photothermal stimulation, there is an advantage that both cell signal measurement, electric stimulation and photo stimulation are possible. In order to induce such a photothermal effect, a technology of using nano metal particles such as gold nano particles or gold nano rods or using a polymer polymer has been developed.

On the other hand, with respect to a method for regulating neuronal activity through photothermal stimulation, Korean Laid-Open Patent Publication No. 10-2015-0047247 (hereinafter abbreviated as 'prior art') includes irradiating a laser to a neuron to which nanoparticles are attached. A method of regulating neuronal activity is disclosed.

In the prior art, the use of nanoparticles requires a combination of particle synthesis, agglomeration prevention surface coating, and chip surface coating technology, and a process of optimizing them for chips is required.

On the other hand, nanomaterials can be divided into types, such as nano thin film, nanoparticles, nanostructures. Nano-film accounts for 67.4% of the world's nano-material consumption (as of 2013), and the proportion of nano-film in the nano-material market is expected to increase to 70.5% in 2019. As a result, it has been possible to devise a microelectrode chip using a nano thin film material which is easy to apply and expected to increase demand in the nano material market, and a method of controlling neuronal activity using the same.

Korean Patent Publication No. 10-2015-0047247

Disclosure of Invention An object of the present invention is to provide a microelectrode chip in which a complicated process for coating a nanomaterial on a microelectrode chip is omitted and a method for controlling neuronal activity using the same.

It is also an object of the present invention to provide a microelectrode chip capable of locally inhibiting neuronal activity and a method for controlling neuronal activity using the same.

Microelectrode chip according to an embodiment of the present invention, the substrate; An electrode pad disposed on a portion of the substrate; An electrode disposed on a portion of the substrate and connected to the electrode pad; An insulating layer disposed on a portion of the substrate where the electrode is not disposed; And a metal thin film disposed on the insulating layer.

In addition, the metal thin film may include gold (AU).

In addition, the metal thin film may have a thickness of 4 to 6nm.

In addition, the microelectrode chip according to the embodiment of the present invention may further include a cell-compatible polymer layer disposed on the metal thin film.

Neuro cell activity control method according to an embodiment of the present invention, preparing a microelectrode chip comprising a metal thin film; Disposing a nerve cell on the microelectrode chip; And irradiating a laser to the microelectrode chip.

In addition, the method for controlling neuronal activity according to an embodiment of the present invention may further include disposing a cell-friendly polymer layer on the microelectrode chip after preparing the microelectrode chip including the metal thin film. Can be.

In addition, preparing the microelectrode chip including the metal thin film may include applying the metal thin film to a substrate by a vacuum deposition method.

In addition, the laser irradiation on the microelectrode chip may be a wavelength of 500 to 900 nm.

The present invention can induce a photothermal effect by irradiating a near-infrared laser to the metal thin film, thereby locally inhibiting the activity of nerve cells disposed on the microelectrode chip. Accordingly, there is an advantage that can be applied to the study of the relationship between neuronal cell activity such as epilepsy and disease through the in vitro culture of neurons and to find the treatment.

In addition, the microelectrode chip according to the present invention has a wider wavelength of light absorbed by the laser to the metal thin film, than the conventional nanoparticles or nanomaterials such as nanostructures.

In addition, the present invention by using a vacuum deposition method by applying a metal thin film of a nano-controlled thickness on the substrate, it is possible to manufacture a microelectrode chip in a simple method. Through this, chip production companies such as the electronics and medical engineering industries can manufacture microelectrode chips at low cost, and productivity can be improved.

1 is a microelectrode chip according to an embodiment of the present invention.
2 is a side cross-sectional view of the microelectrode chip according to the embodiment of the present invention.
3 is a step diagram of a method for controlling neuronal activity according to an embodiment of the present invention.
Figure 4 shows the arrangement of the microelectrode chip, the cell affinity polymer layer and neurons.
5 shows a laser irradiation step and cell activity observation process according to an embodiment of the present invention.
6 is a microelectrode chip in which nerve cells are cultured according to an embodiment of the present invention.
7 is a result of observing the surface of the microelectrode chip coated with a metal thin film according to an embodiment of the present invention.
8 is an absorption spectrum of a microelectrode chip and a metal thin film coated with a metal thin film according to an embodiment of the present invention.
9 is a graph showing a temperature change of a metal thin film when near-infrared laser irradiation.
10 is a result of measuring impedance and phase of the microelectrode chip according to the embodiment of the present invention.
Figure 11 shows the activity ratio of neurons in the near infrared laser irradiation.
12 shows the results of the degree of inhibition of photothermally stimulated neurons.
Figure 13 shows the results of the degree of inhibition of photothermally stimulated neurons.

Hereinafter, with reference to the contents described in the accompanying drawings will be described in detail the present invention. However, the present invention is not limited or limited by the exemplary embodiments. Like reference numerals in the drawings denote members that perform substantially the same function.

The objects and effects of the present invention may be naturally understood or more apparent from the following description, and the objects and effects of the present invention are not limited only by the following description. In addition, in describing the present invention, when it is determined that the detailed description of the known technology related to the present invention may unnecessarily obscure the subject matter of the present invention, the detailed description thereof will be omitted.

1 is a microelectrode chip 1 according to an embodiment of the present invention. 2 is a side cross-sectional view of the microelectrode chip 1 according to the embodiment of the present invention. 1 and 2, the microelectrode chip 1 according to the embodiment of the present invention is disposed on a portion of the electrode pad 113, a portion of the substrate 11, and an electrode pad ( An electrode 111 connected to 113, an insulating layer 17 disposed on a portion where the electrode 111 on the substrate 11 is not disposed, and a metal thin film 13 disposed on the insulating layer 17 are included. In addition, the microelectrode chip 1 may include a polymer layer 15 disposed on the metal thin film 13.

The substrate 11 may include an electrode 111 and an electrode pad 113.

In this embodiment, the substrate 11 may mean any kind of substrate capable of culturing cells, and specifically, may be supplied with a material such as glass, plastic, metal, silicon, PDMS, or the like. The substrate 11 may be provided as a transparent material for the laser to pass through. However, the material provided to the substrate 11 is not limited to the above-described material. The laser described above refers to one type of light source that can be irradiated onto a substrate, and a light source having a different shape from that of the laser may be irradiated. The lasers to be described below are preferably understood in this same context.

In the present embodiment, the electrode 111 may be disposed on a portion of the substrate 11 and connected to the electrode pad 113. The electrode 111 may be for measuring the activity of the neuron 3 during laser irradiation. To this end, the electrode 111 may be provided in the form of points, lines, faces or volumes, and may be provided in a combined configuration thereof. In particular, it can be confirmed that the activity of the neuron 3 is suppressed by confirming that the signal of the neuron measured through the electrode 111 according to the present embodiment is reduced.

Since the electrode 111 is configured to measure the activity of the neuron 3, the electrode 111 should be positioned on the upper surface of the substrate 11, and a plurality of electrodes may be distributed on the substrate 11. Distribution of the plurality of electrodes 111 on the substrate 11 may have a predetermined rule, which may be understood as patterning of the electrodes 111. Patterning methods for electrode 111 include nanoimprint lithography, interference lithography, self-assembled copolymer pattern transfer, electron beam lithography, focused ion beam milling, photolithography, reactive ion etching, wet-etching. Or the like, but is not limited thereto.

The insulating layer 17 may not be formed on the upper surface of the electrode 111, and the insulating layer 17 may be formed on the surface of the microelectrode chip 1 except for the electrode. Accordingly, the surface on which the insulating layer 17 is formed and the surface on which the electrode 111 is distributed have a height difference. The electrode 111 should be provided at a height lower than the height of the insulating layer 17, and even at this height difference should be provided at a height sufficient to measure the activity of the neuron (3).

Electrode 111 is platinum (Pt), palladium (Pd), gold (Au), silver (Ag), titanium (Ti), chromium (Cr), aluminum (Al), copper (Cu), tin (Sn), Molybdenum (Mo), ruthenium (Ru) and indium (In) may optionally include one or more. The active potential of the nerve cell delivered to the electrode 111 may be delivered to the electrode pad 113 and sent out to the outside of the microelectrode chip 1.

In the present embodiment, the electrode pad 113 may be disposed on a portion of the substrate 11. The electrode pad 113 is connected to the electrode 111 may be understood as a configuration for connecting the electrode with the outside. The electrode pad 113 may be connected to the singular or plural electrodes 111. In addition, the substrate 11 may further include a conductive member connecting the electrode pad 113 and the electrode 111. The electrode 111, the electrode pad 113, and the conductive member may be formed of the same material, and may be simultaneously formed by the same process. Therefore, the material of the electrode pad 113 is platinum (Pt), palladium (Pd), gold (Au), silver (Ag), titanium (Ti), chromium (Cr), aluminum (Al), copper (Cu), Tin (Sn), molybdenum (Mo), ruthenium (Ru) and indium (In) may optionally include one or more.

The insulating layer 17 may be disposed at a portion where the electrode 111 on the substrate 11 is not disposed.

In the present embodiment, the insulating layer 17 may electrically separate the substrate 11 and the metal thin film 13. The insulating layer 17 may be disposed at a portion where the electrode is not disposed to form an uneven structure on the substrate. The insulating layer 17 is made of silicon nitride, silicon oxide, aluminum oxide (Al ₂ O ₃ ), bismuth-zinc-niobium oxide, titanium oxide, hafnium oxide, zirconium oxide, tantalum oxide And lanthanum oxide may include one or more selected from the group consisting of.

The height of the insulating layer 17 may be higher than the height of the electrode 111. Through this, insulation between the distributed electrodes 111 can be made easier.

The metal thin film 13 may be disposed on the insulating layer 17.

In the present embodiment, the metal thin film 13 includes an electrode 111 and an insulating layer 17 disposed thereon. It can be applied on the substrate (11). The metal thin film 13 may be applied by vacuum deposition. The metal thin film 13 may be excited with plasmon on the surface of the thin film during near-infrared irradiation, and the excited surface plasmon resonance (SPR) energy may be radioactive damping. Radiation damping generates heat energy, which can be used to inhibit neuronal activity. The metal thin film 13 has less heat generation due to near-infrared absorption than conventional nanomaterials, but has a wide range of wavelengths of absorbed light.

In the microelectrode chip 1 according to the embodiment of the present invention, the metal thin film 13 may include gold (Au).

In the present embodiment, the metal thin film 13 may require a collective vibration of electrons occurring at the metal surface to induce surface plasmon resonance. Metals exhibiting this phenomenon are mainly used for the emission of electrons by external stimulus such as gold, silver, copper, aluminum, etc. and metals having negative dielectric constant, among which silver has the sharpest resonance peak value and excellent surface stability. Gold is commonly used. Accordingly, the metal thin film 13 may be made of metals such as gold, silver, copper, and aluminum, and a metal used in the metal thin film 13 may be selectively used in some of the above-described metals according to a user's needs. .

In the microelectrode chip 1 according to the embodiment of the present invention, the metal thin film 13 may be formed to a thickness of 4 to 6nm.

In this embodiment, the metal thin film 13 may be formed through a vacuum deposition method. In the vacuum deposition method, the thickness of the thin film can be adjusted by several nano units. Accordingly, the user can control the thickness of the metal thin film 13. However, in order to realize the object and effect of the present invention, the metal thin film 13 should have a condition capable of inducing a surface plasmon phenomenon and should be limited to a thickness of high efficiency without disturbing the measurement of the active potential of nerve cells. The range of thickness was limited to the range of 4-6 nm. Related information is supported by the following <Example 1> to <Example 6>.

In the microelectrode chip 1 according to the embodiment of the present invention, the cell-compatible polymer layer 15 disposed on the metal thin film 13 may be further included.

In this embodiment, the cell-friendly polymer layer 15 may be disposed between the nerve cell 3 and the metal thin film 13 when the activity of the nerve cell 3 is measured. Since the cell-friendly polymer layer 15 has cell affinity, the stability of the nerve cell 3 can be improved. The cell-friendly polymer layer 15 may be provided with a polymer material such as polylysine, polydopamine, laminin, fibronectin, or the like.

Hereinafter, according to the embodiment of the present invention A method of controlling neuronal activity using the microelectrode chip 1 will be described. In the following description, overlapping parts related to the configuration and characteristics of the microelectrode chip 1 will be omitted, and a description related to the method for controlling the activity of neurons will be described.

3 is a step diagram of a method for controlling neuronal activity according to an embodiment of the present invention. 4 is a diagram illustrating a method of controlling neuronal activity according to an embodiment of the present invention. Referring to FIG. 3, the method for controlling neuronal activity according to the present embodiment includes preparing a microelectrode chip including a metal thin film (S1), arranging neurons on the microelectrode chip (S5), and a microelectrode. Irradiating a laser onto the chip (S7).

In preparing the microelectrode chip including the metal thin film (S1), the microelectrode chip 1 may be the microelectrode chip 1 according to the embodiment of the present invention described above. In this step, the metal thin film 13 may be formed by a vacuum deposition method.

Next, disposing a nerve cell on the microelectrode chip (S5) can be performed. The neuron 3 may be selected from the group consisting of microorganisms, hippocampal neurons, sensory neurons, associated neurons and motor neurons of animal and plant cells, but is not limited thereto.

Next, the laser is irradiated to the microelectrode chip in which the nerve cells are arranged. 5 shows a laser irradiation step and cell activity observation process according to an embodiment of the present invention. Referring to FIG. 5, the laser irradiation step (S7) may irradiate a laser to the microelectrode chip 1 by selecting a wavelength.

In this embodiment, the laser can be irradiated through a controllable medium. The control medium may include any medium capable of executing stimulus generating software, such as a personal computer or laptop. The laser irradiation command can then be delivered to the stimulator via stimulus generation software and the laser irradiation signal from the stimulator to the near infrared controller. The laser is irradiated through the near infrared controller, and may be irradiated in a direction passing from the lower surface of the substrate to the upper surface.

In the laser irradiation, the photothermal effect may be induced through the surface plasmon resonance in the metal thin film 13. Accordingly, the nerve cells 3 disposed on the substrate 11 may be stimulated, and the action potential of the stimulated nerve cells 3 may be changed. The changed action potential is input through the electrode 111 and amplified by the potential through the signal amplifier, and data collection and output may be performed. The laser can be irradiated to an extent that can sufficiently cover the entire area where the nerve cells 3 are cultured. In addition, the irradiation direction of the laser can be irradiated to be perpendicular to the surface on which the nerve cells 3 are cultured.

The laser irradiated in the laser irradiation step S7 may be a wavelength of 500 to 900 nm. In the present embodiment, the metal thin film 13 may absorb a wider wavelength than conventional materials such as nanoparticles, nanorods, and nanostructures. Therefore, the user can selectively irradiate the wavelength of the near infrared band within the above-described range.

In a method of controlling neuronal activity according to another embodiment of the present invention, after preparing a microelectrode chip including a metal thin film (S1), disposing a cell-friendly polymer layer on the microelectrode chip (S3). It may further include.

The cell-friendly polymer layer 15 may be disposed between the nerve cell 3 and the metal thin film 13 when the activity of the nerve cell 3 is measured. Through this, it is possible to improve the stability of the neuron (3). The cell-friendly polymer layer 15 may be provided with a polymer material such as polylysine, polydopamine, laminin, fibronectin, and the like.

The formation of the polymer layer 15 plays a role of allowing the nerve cells 3 to stably grow, and is an essential process even when culturing the nerve cells on the microelectrode chip 11 except the metal thin film 13. Can be understood. That is, the formation of the polymer layer 15 may be understood as a process that is always used in the culture of nerve cells for experiments other than the cell culture for photothermal stimulation.

< Example 1 > Nerve cells distributed Micro electrode chip

6 is according to an embodiment of the present invention It shows that the nerve cells are cultured on the microelectrode chip. Referring to FIG. 6, it can be seen that the neurons are uniformly distributed on the microelectrode chip coated with the metal thin film containing gold. The distributed neurons survive to the time period of the signal, thereby measuring the activity of the neurons at each electrode.

< Example 2 > Formed with different thickness Metal thin film  Surface comparison

7 is a result of observing the surface of the microelectrode chip according to the embodiment of the present invention. AFM (Atomic-force microscopy) was used for the observation. Referring to FIG. 7, the result disclosed on the left is a result of observing a metal thin film having a thickness of 2 nm, and the result disclosed on the right is a result of observing a metal thin film having a thickness of 5 nm. In addition, the results disclosed at the bottom of the figure observed a 20 x 20 um area, the results disclosed at the top is a result of observing the 1 x 1 um area. As a result, it can be seen that the particles on the surface of the metal thin film having a thickness of 5 nm is larger than the metal thin film having a thickness of 2 nm. In addition, it can be seen that both samples evenly formed on the surface of the metal thin film.

< Example 3 > With different thickness Metal film  Formed Of micro electrode chip  Absorption Spectrum Comparison

8 is according to an embodiment of the present invention Absorption spectra of microelectrode chips and metal thin films. 8 (a) and 8 (b) disclose microelectrode chips in which metal thin films are manufactured in different thicknesses. 8 (a) shows a 2 nm metal thin film, and FIG. 8 (b) shows a micro electrode chip on which a 5 nm metal thin film is made. The thickness of the metal thin film applied to the microelectrode chip used as the sample is 2 nm, 5 nm and 7 nm. Then, the absorption wavelength was confirmed using a UV-vis spectrometer on each of the manufactured microelectrode chips.

In the results of this example, a 7 nm thick metal thin film exhibited the highest absorption, but a short circuit phenomenon was found in which the signal could not be measured in the microelectrode chip. That is, the metal thin film is formed thinner than the thickness of 7nm it can be confirmed that the conditions for preventing the short-circuit phenomenon. Therefore, all the experimental examples below compared the data limited to the thickness of the metal thin film in the range less than 7nm, in particular compared the characteristics of the metal thin film having a thickness of 2nm and 5nm.

FIG. 8 (c) shows a graph comparing the results of the metal thin films of 2 nm and 5 nm thicknesses in which the measurement was made stable. Referring to the graph, the optical density of the 5 nm thick metal thin film was measured for better efficiency in all near infrared wavelength ranges.

< Example 4 Laser irradiation Metal thin film  Temperature change comparison

9 is a graph showing the temperature change of the metal thin film when the near infrared laser irradiation. For the temperature change, the temperature of the metal thin film was measured for 3 minutes in the air by using an IR camera, and the change amount was quantified by subtracting the average value of the measured values for 2 minutes before / after near infrared irradiation. The lasers used are divided into intensities of 3, 6, 9, 12, 15 mW / mm ² .

The graph disclosed at the top of FIG. 9 shows the temperature change of the 2 nm thick metal thin film, and the graph disclosed at the bottom shows the temperature change of the 5 nm thick metal thin film. Comparing the two graphs, it can be seen that the metal thin film having a thickness of 5 nm generates more heat than the metal thin film having a thickness of 2 nm for all near infrared intensities. In particular, it can be seen that a 5 nm thick metal film detects a temperature change of up to 25 degrees in air, while a 2 nm thick metal film detects a temperature change of less than 20 degrees.

< Example 5 > Metal film  Formed Of microfluidic chips  Impedance and Phase  Measure

10 is a result of measuring impedance and phase of the microelectrode chip according to the embodiment of the present invention. The left side of FIG. 10 shows the impedance measurement graph of a metal thin film, and the right side shows the graph of a phase measurement result.

In the case of impedance measurement, the impedance of each thickness was measured to measure cellular activity. In particular, it can be seen that the metal thin film having a thickness of 5 nm is measured higher than the metal thin film having a thickness of 2 nm.

In the case of phase measurement, neuronal activity was determined at two thickness levels.

< Example 6 > Measure signal of nerve cell activity

Figure 11 shows the activity ratio of neurons in the near infrared laser irradiation. This example shows the result of a state in which an active signal of a neuron is not processed.

Referring to FIG. 11, in an experiment not irradiated with near infrared rays, it can be seen that the activation signal of neurons is continuously generated. On the other hand, in the experiments irradiated with near-infrared, it can be seen that the activity of neurons is suppressed during the irradiation. In addition, the results of neuron observation after the experiment with near-infrared light showed that the activity was not affected and was continuously maintained in the living state.

12 shows the results of the degree of inhibition of photothermally stimulated neurons. In this example, the measured signals were processed and analyzed quantitatively by Peristimulus time histogram (PSTF). The experiment was conducted with a near-infrared stimulation experiment in a period of 14 days to 21 days after the culture in which the action potential of neurons is actively induced.

Referring to FIG. 12, the vertical axis of the graph means spikes / sec (number of signals / second) and the horizontal axis means time (sec). During the experiment, when the activity of nerve cells in the electrode of the microelectrode chip is measured, the dots are continuously spotted.

In particular, -10 to 0 seconds, 10 to 20 seconds in the two graphs on the right side is the state without the laser stimulation, 0 to 10 seconds is the state of the laser stimulation. In addition, in every graph, a small dot appearing at the top of the graph between -10 and 20 seconds is the number of signals measured in the microelectrode chip at that time, and a single row formed by each point represents the number of attempts. The result is 30 attempts, with 30 rows.

In particular, in the case of a microelectrode chip having a 5 nm thick gold nano thin film, it is confirmed that the activity of the neuron is significantly reduced during the 0 to 10 seconds when the near infrared laser is irradiated, so that almost no spots are photographed.

Figure 13 shows the results of the degree of inhibition of photothermally stimulated neurons. In this example, the degree of inhibition of neuronal activity to the intensity of photothermal stimulation was measured according to the thickness of the metal thin film. The lasers irradiated are 3, 6, 9, 12, 15, 18 and 21 mW / mm ² It was irradiated for 10 seconds at the intensity of, and the experiment was repeated a total of 30 times without irradiating near infrared rays for 20 seconds. As a control of the experiment, the activity potential inhibition of neurons was quantified by comparing the activity potential measured at 0 mW / mm ² for 15 minutes.

As a result, in the microelectrode chip sample in which the metal thin film of 2 nm thickness was formed, a burst signal was generated in some signals, but it was confirmed that it was easily suppressed during photostimulation. As shown in the graph of FIG. 13, it can be seen that the activity of nerve cells gradually decreases as the near infrared intensity increases in both the microelectrode chips having the metal thin films of 2 nm and 5 nm thickness. In particular, in the microelectrode chip having a metal thin film of 5 nm, it was confirmed that the activity of neurons was significantly suppressed even in the near-infrared ray of low intensity than the chip having a thickness of 2 nm. Through this, it was confirmed that the microelectrode chip in which the metal thin film was formed with a thickness of 5 nm was a sample suitable for inhibiting the activity of neurons.

Although the present invention has been described in detail through the representative embodiments above, those skilled in the art to which the present invention pertains will appreciate that various modifications can be made without departing from the scope of the present invention. will be. Therefore, the scope of the present invention should not be limited to the described embodiments, but should be determined by all changes or modifications derived from the claims and the equivalent concepts as well as the following claims.

1: micro electrode chip
11: substrate
111: electrode
113: electrode pad
13: metal thin film
15: polymer layer
17: insulation layer
3: nerve cells
S1: microelectrode chip preparation step
S3: polymer layer placement step
S5: neuronal cell placement
S7: laser irradiation step

Claims (8)

Board;
An electrode pad disposed on a portion of the substrate;
An electrode disposed on a portion of the substrate and connected to the electrode pad;
An insulating layer disposed on a portion of the substrate where the electrode is not disposed; And
And a metal thin film disposed on the insulating layer.
The method of claim 1,
The metal thin film,
Microelectrode chip containing gold (AU).
The method of claim 1,
The metal thin film,
Microelectrode chip having a thickness of 4 to 6nm.
The method of claim 1,
The microelectrode chip further comprises a cell-compatible polymer layer disposed on the metal thin film.
Preparing a microelectrode chip;
Disposing a nerve cell on the microelectrode chip; And
Irradiating a laser on the microelectrode chip; including;
The micro electrode chip,
Board; An electrode pad disposed on a portion of the substrate; An electrode disposed on a portion of the substrate and connected to the electrode pad; An insulating layer disposed on a portion of the substrate where the electrode is not disposed; And a metal thin film disposed on the insulating layer.
The method of claim 5,
After preparing the microelectrode chip including the metal thin film, neuron activity control method further comprising the step of disposing a cell-compatible polymer layer on the microelectrode chip.
The method of claim 5,
The preparing of the microelectrode chip including the metal thin film may include applying the metal thin film to a substrate by a vacuum deposition method.
The method of claim 5,
The laser irradiation on the microelectrode chip has a wavelength of 500 to 900 nm nerve cell activity control method.

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